Team:TCU Taiwan/Notebook

From 2014.igem.org

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<li><a href="#" style="cursor:default"><font size="5">Team</font></a>
<li><a href="#" style="cursor:default"><font size="5">Team</font></a>
     <ul>
     <ul>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Team Members</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Members</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Achievements"><font size="3">Achievements</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
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   <li><a href="#" style="cursor:default"><font size="5">Project</font></a>
   <li><a href="#" style="cursor:default"><font size="5">Project</font></a>
     <ul>
     <ul>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Project"><font size="3">Project</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Project"><font size="3">Introduction</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Parts"><font size="3">Parts</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Parts"><font size="3">Parts</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/SystemDesign"><font size="3">System Design</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/M13Phage"><font size="3">M13 Phage</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/CRISPR"><font size="3">CRISPR</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/FutureWorks"><font size="3">Future Works</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/HumanPratics"><font size="3">Human Pratics</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/HumanPratics"><font size="3">Human Pratics</font></a></li>
     </ul>
     </ul>
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</td>
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<tr><td colspan="3" height="53" background="#F3E4C2"><img src="https://static.igem.org/mediawiki/2014/c/c1/TCU-Line.png" width="100%"></td></tr>
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<tr><td colspan="3"background="#F3E4C2"><img src="https://static.igem.org/mediawiki/2014/c/c1/TCU-Line.png" width="100%"></td></tr>
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  <td colspan="3"  height="15px"></td>
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<tr> <td colspan="3"  height="15px" style="background-color:#FFF2B5"><font face="Trebuchet MS" size="7" color="#A66B38">iGEM materials</font></td></tr>
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<tr><td bgColor="#ffffff" colspan="3" height="10px"> </tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Bacteria medium</font><br>
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  </legend>
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<div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*LB(Luria-Bertani) broth: 1% Trytone, 1% NaCl, 0.5%  Yeast extract in the ddH2O<br>
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*LB  agar plate:LB broth add 1.5% agar.If you want to add selection marker example  for ampicillin or chloramphenicol. To culture <strong><em>E.coli</em></strong>, ampicillin is  added 100 mg/ml  for usage, chloramphenicol is added 30 mg/ml for usage,  kanamycin is added 70 mg/ml for usage</p>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Plasmid isolation</font><br></legend>
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  <div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*Solution  I: 50 mM glucose、25 mM  Tris-HCl (pH 8.0)、10 mM EDTA<br>
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*Solution II:0.2N NaOH and 1% SDS(Sodium dodecyl sulfate&nbsp;)  mix<br>
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*Solution III: 5 M  potassium acetate (pH4.8), 11.5% glacial acetic acid<br>
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*RNase A(10 mg/ml)<br>
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*Phenol chloroform<br>
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*Chloroform<br>
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*95%  Ethanol<br>
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*Tris-EDTA(TE)buffer:10  mM Tris-Cl(pH 7.5) 1 mM EDTA<br>
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*pBluescript  SK(-)<br>
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*BBa_K1218011<br>
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*BBa_K914003<br>
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*BBa_I13521</p>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Quick screening</font><br></legend>
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<div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*Quick  screening buffer(Tris-Cl pH 12.6, 3% SDS)<br>
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*Phenol  chloroform<br>
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*Chloroform</p>
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</font>
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</div>
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</tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px">
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</tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Enzyme digestion</font><br>
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</legend>
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<div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*T4  ligase<br>
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*10X  ligation buffer<br>
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*<em>Eco</em>RI<br>
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*<em>Pst</em>I<br>
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*<em>Spe</em>I<br>
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*<em>Xba</em>I<br>
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*BSA  0.1%</p>
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</font>
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</div>
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</tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px">
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</tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Bacteria strains and others</font><br></legend>
 +
<div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*DH5a<br>
 +
    *JM101<br>
 +
    *0.8%  and 2% agarose gel<br>
 +
    *95%  Ethanol<br>
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    *6X  loading dye<br>
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    *1.5  ml centrifuge tube<br>
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    *250  ml flask<br>
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    *M13KO7(1*1011  pfu/ml)from <em>NEW ENGLAND BioLabs</em></p></font>
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</div>
 +
    </fieldset>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px">
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</tr>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Polymerase chain reaction</font><br></legend>
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<div class="wrapper" style="background-color: white;">
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  <font size="3" face="Verdana" color="#333"><p>*<em>Top</em> DNA polymerase<br>
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*dNTP(dATP,  dCTP, dGTP, dTTP) 250 mM<br>
 +
*Tris-HCl(pH9.0)  10 mM<br>
 +
*KCl  30 mM<br>
 +
*MgCl2  1.5 mM</p>
 +
</font>
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</div>
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<tr>
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  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
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  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#82C0AF">Instruments</font><br></legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*VCM-620, laminal flow<br>
 +
*Biofuge pico, Heraeus<br>
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* OS1500-H, TSK Oribital Shaking Iucubator<br>
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* Biowave<br>
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* Vortex-GENE 2<br>
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* Autoclave AS-1060L<br>
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* GenAmp PCR System 2400, Applied  Biosystems<br>
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*BU-420,  YIHDER<br>
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*5804R,  eppendorf Centrifuge</p>
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<tr> <td colspan="3"  height="5px"> </td></tr>
<!--requirements section -->
<!--requirements section -->
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<tr><td colspan="3"> <h3>Notebook</h3></td></tr>
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<tr><td colspan="3" style="background-color:#FFF2B5"><font face="Trebuchet MS" size="7" color="#A66B38">Life Records</font></td></tr>
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  <td width="45%"  valign="top" height="30">&nbsp;</td>
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<p>You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well. </p>
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  <td  valign="top">&nbsp;</td>
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   <td>&nbsp;</td>
   <td>&nbsp;</td>
   <td></td>
   <td></td>
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    <td height="50px">&nbsp;</td>
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     <div style="width:70%; height:1250px">
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    <div id="accordion">
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        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 1</font><font face="Trebuchet MS" size="6" color="#90B849"> 6/22~6/28</font></h6>
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        <div align="left">
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          <p><h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5><br>
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          <font face="Trebuchet MS" size="4" color="#333">Prepare for the competent cell.</font>
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          <p><font face="Trebuchet MS" size="4" color="#333">BoBo and Ted decided to multiply plasmid within bacteria but not PCR, like this:<br>
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          Transformation ( Cas9 in pSB1C3 into DH5α)<br>Incubate DH5α with pSB1C3</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Extract plasmid with pSB1C3 from DH5α.</font>          </p>
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          <p><h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5><br>
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          <font face="Trebuchet MS" size="4" color="#333">Prepare Human practice (ppt).<br>
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          Practice presentation.<br>
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          Design gRNA for AmpR.<br>
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          Discuss about project. Project Name is E.Maat for now (came up by Sharon).<br>
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          Order restraction enzyme(PstI、EcoRI、XbaI、SpeI). </font></p>
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        </div>
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        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 2</font><font face="Trebuchet MS" size="6" color="#90B849"> 6/29~7/5</font></h6>     
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        <div align="left">
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
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          <br>
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          <font face="Trebuchet MS" size="4" color="#333">Successfully transform biobrick BBa_I13521、BBa_ K914003、BBa_ K1218011 into DH5α (plasmid: pSB1C3<br>Extract pSB1C3 after incubation<br>Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 is successfully extracted<br>Prepared Cm. plate、LB. plate<br>
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Prepared competent cell
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</font></p>
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          <p>&nbsp;</p>
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
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          <br>
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          <p><font face="Trebuchet MS" size="4" color="#333">Get foundation from nearby store (yeah!!!!!)<br>
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            Decided to have iGEM contest in local school as human practice</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Others don’t think E.Maat is a good name, under re-consideration </font></p>
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        </div>
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        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 3</font><font face="Trebuchet MS" size="6" color="#90B849"> 7/6~7/12</font></h6>
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        <div align="left">
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Extract plasmid pSB1C3 with BBa_I13521、BBa_ K914003、BBa_ K1218011</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Prepare Cm. plate、LB. plate </font>
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            </p>
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          </p>
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          <p>         
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Something wrong with gRNA for AmpR, redesign</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Second project theme came out by Denise and Regina: Trogene horse. Needs later discuss to decided which one as theme( E.Maat or Trogene Horse)</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Try to get foundation from Biotechnology company on e-mail.</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Prepare for igem conference in NCTU-Formosa (ppt) </font></p>
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          </p>
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        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 4</font><font face="Trebuchet MS" size="6" color="#90B849"> 7/13~7/19</font></h6>
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        <div align="left">
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Incubate E.coli DH5α with BBa_I13521、BBa_ K914003、BBa_ K1218011(in pSB1C3 )</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Extract pSB1C3 BBa_I13521、BBa_ K914003、BBa_ K1218011 </font>
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            </p>
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          </p>
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          <p>         
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Final Theme decided: Trogene Horse</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Prepare for igem conference in NCTU-Formosa (ppt)</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333"> Designing team logo and project logo</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">House started working on wiki</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Decided to have a synthetic biology contest in National Hualien Girls’ Senior High School </font></p>
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          </p>
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      <h6 align="left"> <font face="Trebuchet MS" size="6" color="#A66B38">Week 5</font><font face="Trebuchet MS" size="6" color="#90B849"> 7/20~7/26</font></h6>
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        <div align="left">
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Incubate:</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521、BBa_ K914003、BBa_ K1218011(in pSB1C3 )</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">2. E.coli top10 with pBluescript II SK (-)</font></p>
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          <p>&nbsp;</p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521、BBa_ K914003、BBa_ K1218011</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">2. pBluescript</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted. </font>
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            </p>
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          </p>
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          <p>         
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
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          <p><br>
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            <font face="Trebuchet MS" size="4" color="#333">Prepare for igem conference in NCTU-Formosa (ppt)
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            </font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333"> Designing team logo and project logo</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">House working on wiki construction</font></p>
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          <p><font face="Trebuchet MS" size="4" color="#333">Prepare for the contest in National Hualien Girls’ Senior High School (ppt)</font></p>
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          </p>
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        </div>
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        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 6</font><font face="Trebuchet MS" size="6" color="#90B849"> 7/27~8/2</font></h6>
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        <div align="left">
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          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Incubate:</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521、BBa_ K914003、BBa_ K1218011(in pSB1C3 )</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. E.coli top10 with pBluescript II SK (-)</font></p>
+
-
          <p>&nbsp;</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521、BBa_ K914003、BBa_ K1218011</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. pBluescript</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted. </font>
+
-
            </p>
+
-
          </p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Prepare for igem conference in NCTU-Formosa (ppt).
+
-
            </font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333"> Have a synthetic biology contest in National Hualien Girls’ Senior High School.</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Order primer for gRNA
+
-
            team logo and project logo confirmed. </font></p>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 7</font><font face="Trebuchet MS" size="6" color="#90B849"> 8/3~8/7</font></h6>
+
-
        <div align="left">
+
-
          <h5>
+
-
          <p><font face="Trebuchet MS" size="4" color="#E72B33">Attending iGEM conference in NCTU-Formosa </font>
+
-
            </p>
+
-
          </p>
+
-
          <p><img src="https://static.igem.org/mediawiki/2014/4/47/1621918_793376414029197_3582457878420185460_n.jpg" width="60%"></p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 8</font><font face="Trebuchet MS" size="6" color="#90B849"> 8/10~16</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Incubate:</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521、BBa_ K914003、BBa_ K1218011(in pSB1C3 )</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. E.coli top10 with pBluescript II SK (-)</font></p>
+
-
          <p>&nbsp;</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521、BBa_ K914003、BBa_ K1218011</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. pBluescript</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font>.</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Purification of RFP sequence from pSB1C3
+
-
            Prepare Cm. plate、LB. plate</font></p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <br>
+
-
          <font face="Trebuchet MS" size="4" color="#333">Team uniform under design. </font>
+
-
         
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 9</font><font face="Trebuchet MS" size="6" color="#90B849"> 8/17~8/23</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Incubate:</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521、BBa_ K914003、BBa_ K1218011(in pSB1C3 )</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. E.coli top10 with pBluescript II SK (-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521、BBa_ K914003、BBa_ K1218011</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. pBluescript</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted.</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Purification of RFP sequence from pSB1C3. </font>
+
-
            </p>
+
-
         
+
-
         
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 10</font><font face="Trebuchet MS" size="6" color="#90B849"> 8/24~8/30</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Ligation of RFP and pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Transformation clone RFP/pBluescript SK(-) to DH5α</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333"> Extract plasmid clone RFP/pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted. </font>
+
-
            </p>
+
-
          </p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Design badge as small gift</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Order Helper phage for pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Make movie for project introduction of Trogene horse</font>.</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Prepare iGEM presentation ppt、poster. </font></p>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 11</font><font face="Trebuchet MS" size="6" color="#90B849"> 8/31~9/6</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Incubate:</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521、(in pSB1C3 )</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. E.coli top10 with pBluescript II SK (-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">2. pBluescript</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Purification of RFP sequence from pSB1C3</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Ligation Clone RFP/pBluescript SK(-)to DH5α
+
-
            Store BBa_I13521、BBa_ K914003、BBa_ K1218011、clone RFP/pBluescript SK(-) </font>
+
-
            </p>
+
-
          </p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Make movie for project introduction of Trogene</font>.</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333"> horse
+
-
            Prepare iGEM presentation ppt、poster. </font></p>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 12</font><font face="Trebuchet MS" size="6" color="#90B849"> 9/7~9/13</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Ligation Clone RFP/pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract plasmid clone RFP/pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Modeling for phage injection (Thanks for NCTU-Formosa’s help)</font></p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <br>
+
-
          <font face="Trebuchet MS" size="4" color="#333">Prepare iGEM presentation ppt、poster. </font>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 13</font><font face="Trebuchet MS" size="6" color="#90B849"> 9/14~9/20</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Ligation Clone RFP/pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extractplasmid clone RFP/pBluescript SK(-)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Prepare competent cell(JM101)</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">incubate:</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. E.coli DH5α with BBa_I13521(pBluescript)、BBa_K1473001~3003(in pSB1C3 )</font></p>
+
-
          <p>&nbsp;</p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract :</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">1. pSB1C3: BBa_I13521、BBa_K1473001~3003</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333"> PCR: tracrRNA、Cas 9+crRNA</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Modeling for phage injection </font>
+
-
            </p>
+
-
          </p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Prepare iGEM presentation ppt、poster</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Wiki construction </font></p>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 14</font><font face="Trebuchet MS" size="6" color="#90B849"> 9/21~9/27</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">experimental part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Ligation gRNA/ pSB1C3</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Transformation gRNA/ pSB1C3 to DH5α</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Incubate gRNA/ pSB1C3 in E.coli JM101</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Extract plasmid gRNA/ pSB1C3 in E.coli JM101</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Use Enzyme digestion(PstI and EcoRI) to confirm whether pSB1C3 and pBluescript is successfully extracted</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">PCR trRNA、Cas 9+crRNA</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Prepare modeling</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">3 new biobrick finished </font>
+
-
            </p>
+
-
          </p>
+
-
          <p>         
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Have a iGEM contest(HP) in Tzu chi university</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Prepare presentation ppt、poster</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Wiki construction </font></p>
+
-
          </p>
+
-
        </div>
+
-
       
+
-
        <h6 align="left"><font face="Trebuchet MS" size="6" color="#A66B38">Week 15</font><font face="Trebuchet MS" size="6" color="#90B849"> 9/28~10/4</font></h6>
+
-
        <div align="left">
+
-
          <h5><font face="Trebuchet MS" size="4" color="#0879A6">other part:</font></h5>
+
-
          <p><br>
+
-
            <font face="Trebuchet MS" size="4" color="#333">Prepare iGEM presentation ppt、poster</font></p>
+
-
          <p><font face="Trebuchet MS" size="4" color="#333">Wiki construction </font></p>
+
-
          </p>
+
-
        </div>
+
-
    </div>
+
-
</div></td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td colspan="3" height="1px" background="#F3E4C2"><img src="https://static.igem.org/mediawiki/2014/c/c1/TCU-Line.png" width="100%"></td>
+
     <td align="center"><iframe src='http://cdn.knightlab.com/libs/timeline/latest/embed/index.html?source=0An3IHGTTAU59dFRRcGpFa0lucU92TzUxZ0hFYWZKaVE&font=Bevan-PotanoSans&maptype=toner&lang=en&height=650' width='100%' height='650' frameborder='0'></iframe></td>
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td><table width="100%" border="0" align="center" style="background-color:#F2DE79" height="150px">
+
     <td align="center" height="30px">&nbsp;</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="center" height="30px"><table width="70%" border="0" cellspacing="0" cellpadding="0">
 +
      <tr>
 +
        <td height="30">&nbsp;</td>
 +
      </tr>
 +
      <tr>
 +
        <td style="background-color:#FFF2B5" align="left"><font face="Trebuchet MS" size="7" color="#A66B38">S<font face="Trebuchet MS" size="7" color="#A66B38">tudy Together</font></font></td>
 +
      </tr>
 +
      <tr>
 +
        <td style="background-color:#FFF2B5" align="left">&nbsp;</td>
 +
      </tr>
 +
      <tr>
 +
        <td><iframe width="720" height="482" frameborder="0" src="http://files.slidemypics.com/app/js/iframe.html?bg_color=fafafa&amp;hash=67635bbf913055c734b13d75a1461392&amp;r=0.02183609106577933"></iframe></td>
 +
      </tr>
 +
      <tr>
 +
        <td height="30">&nbsp;</td>
 +
      </tr>
 +
    </table>
 +
   
 +
    <table width="70%" border="0" cellspacing="0" cellpadding="0">
 +
      <tr>
 +
        <td height="30">&nbsp;</td>
 +
      </tr>
 +
      <tr>
 +
        <td style="background-color:#FFF2B5" align="right"><font face="Trebuchet MS" size="7" color="#A66B38"><font face="Trebuchet MS" size="7" color="#A66B38">Conference in NCTU</font></font></td>
 +
      </tr>
 +
      <tr>
 +
        <td style="background-color:#FFF2B5" align="right">&nbsp;</td>
 +
      </tr>
 +
      <tr>
 +
        <td align="right"><iframe width="720" height="482" frameborder="0" src="http://files.slidemypics.com/app/js/iframe.html?bg_color=ffffff&amp;hash=035d3355e0b6454be558e6d9a1461398&amp;r=0.9810947910882533"></iframe></td>
 +
      </tr>
 +
      <tr>
 +
        <td height="30">&nbsp;</td>
 +
      </tr>
 +
      <tr>
 +
        <td height="30"><table width="100%" border="0" cellspacing="0" cellpadding="0">
 +
 
 +
      <tr>
 +
        <td style="background-color:#FFF2B5" align="left"><font face="Trebuchet MS" size="7" color="#A66B38"><font face="Trebuchet MS" size="7" color="#A66B38">Trip in Chihsingtan Beach</font></font></td>
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        <td style="background-color:#FFF2B5" align="left">&nbsp;</td>
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        <td><iframe width="720" height="482" frameborder="0" src="http://files.slidemypics.com/app/js/iframe.html?bg_color=ffffff&amp;hash=fb44d802e37470fe77b56b41a1461402&amp;r=0.6266446833033115"></iframe></td>
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<table width="100%" border="0" align="center" style="background-color:#F2DE79" height="150px">
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     <td width="15%" align="center"><div id="gotop">^</div>
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    <a href="mailto: tcutaiwan@gmail.com" target="_blank" title="Gmail"><img src="https://static.igem.org/mediawiki/2014/c/c4/TCU%E3%84%A6Gmail.png" width="30"></a>&nbsp;
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      <table width="70%" border="0" cellspacing="0" cellpadding="0" align="center" style="background-color:#F2DE79">
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        <tr align="center">
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Members">Team Members</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Parts">Parts</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Pratics">Human Pratics</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Modeling">Modeling</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Safety">Safety</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Notebook">Notebook</font></td>
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          <td><font size="2" face="Verdana" color="#F2DE79" id="Attributions">Attributions</font></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Team" title="Team Members"><img src="https://static.igem.org/mediawiki/2014/1/1a/TCU-Team-logo.png" width="100"onMouseOver="this.width=this.width*1.1;Members.color='#333'" onMouseOut="this.width=this.width/1.1;Members.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Parts" title="Parts"><img src="https://static.igem.org/mediawiki/2014/e/e5/TCU-Part-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Parts.color='#333'" onMouseOut="this.width=this.width/1.1;Parts.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/HumanPratics" title="Human Pratics"><img src="https://static.igem.org/mediawiki/2014/d/dd/TCU-Human-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Pratics.color='#333'" onMouseOut="this.width=this.width/1.1;Pratics.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Modeling" title="Modeling"><img src="https://static.igem.org/mediawiki/2014/3/3f/TCU-Modeling-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Modeling.color='#333'" onMouseOut="this.width=this.width/1.1;Modeling.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Safety" title="Safety"><img src="https://static.igem.org/mediawiki/2014/2/21/TCU-Safety-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Safety.color='#333'" onMouseOut="this.width=this.width/1.1;Safety.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Notebook" title="Notebook"><img src="https://static.igem.org/mediawiki/2014/2/25/TCU-Notebook-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Notebook.color='#333'" onMouseOut="this.width=this.width/1.1;Notebook.color='#F2DE79'"></a></td>
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          <td><a href="https://2014.igem.org/Team:TCU_Taiwan/Attributions" title="Attributions"><img src="https://static.igem.org/mediawiki/2014/5/51/TCU-Attributions-logo.png" width="100" onMouseOver="this.width=this.width*1.1;Attributions.color='#333'" onMouseOut="this.width=this.width/1.1;Attributions.color='#F2DE79'"></a></td>
 +
        </tr>
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      <p><font size="2" face="Verdana" color="#333" id="tip">Lost the way? Use it to help you if you're lost</font><font color="#333" size="2" face="Verdana">.</font></p>
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     <p><img src="http://i.creativecommons.org/l/by/3.0/88x31.png" width="100"></p>      <p></p></td>
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Latest revision as of 18:19, 17 October 2014

 
Notebook
  
iGEM materials
Bacteria medium

*LB(Luria-Bertani) broth: 1% Trytone, 1% NaCl, 0.5% Yeast extract in the ddH2O
*LB agar plate:LB broth add 1.5% agar.If you want to add selection marker example for ampicillin or chloramphenicol. To culture E.coli, ampicillin is added 100 mg/ml for usage, chloramphenicol is added 30 mg/ml for usage, kanamycin is added 70 mg/ml for usage
Plasmid isolation

*Solution I: 50 mM glucose、25 mM Tris-HCl (pH 8.0)、10 mM EDTA
*Solution II:0.2N NaOH and 1% SDS(Sodium dodecyl sulfate ) mix
*Solution III: 5 M potassium acetate (pH4.8), 11.5% glacial acetic acid
*RNase A(10 mg/ml)
*Phenol chloroform
*Chloroform
*95% Ethanol
*Tris-EDTA(TE)buffer:10 mM Tris-Cl(pH 7.5) 1 mM EDTA
*pBluescript SK(-)
*BBa_K1218011
*BBa_K914003
*BBa_I13521
Quick screening

*Quick screening buffer(Tris-Cl pH 12.6, 3% SDS)
*Phenol chloroform
*Chloroform
Enzyme digestion

*T4 ligase
*10X ligation buffer
*EcoRI
*PstI
*SpeI
*XbaI
*BSA 0.1%
Bacteria strains and others

*DH5a
*JM101
*0.8% and 2% agarose gel
*95% Ethanol
*6X loading dye
*1.5 ml centrifuge tube
*250 ml flask
*M13KO7(1*1011 pfu/ml)from NEW ENGLAND BioLabs
Polymerase chain reaction

*Top DNA polymerase
*dNTP(dATP, dCTP, dGTP, dTTP) 250 mM
*Tris-HCl(pH9.0) 10 mM
*KCl 30 mM
*MgCl2 1.5 mM
Instruments

*VCM-620, laminal flow
*Biofuge pico, Heraeus
* OS1500-H, TSK Oribital Shaking Iucubator
* Biowave
* Vortex-GENE 2
* Autoclave AS-1060L
* GenAmp PCR System 2400, Applied Biosystems
*BU-420, YIHDER
*5804R, eppendorf Centrifuge
Life Records
   
 
 
 
Study Together
 
 
 
Conference in NCTU
 
 
Trip in Chihsingtan Beach
 
 
 
 
^


    
Team Members Project Parts Human Pratics Modeling Safety Notebook Attributions

Lost the way? Use it to help you if you're lost.

^

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